Hara H, Morita M, Iwaki T, Hatae T, Itoyama Y, Kitamoto T, Akizuki S, Goto I, Watanabe T
Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
J Exp Med. 1994 Sep 1;180(3):831-9. doi: 10.1084/jem.180.3.831.
Identification of the localization of human T lymphotrophic virus type I (HTLV-I) proviral DNA in the central nervous system (CNS) is crucial to the understanding of the pathogenesis of HTLV-I-associated myelopathy (HAM)/tropical spastic paraparesis (TSP) pathogenesis. We have developed a sensitive detection method, called two-step polymerase chain reaction (PCR) in situ hybridization, which enabled us to detect the HTLV-I proviral DNA in paraffin-embedded spinal cord tissue sections from HAM/TSP patients. HTLV-I proviral DNA was detected only in the nucleus of lymphocytes that had infiltrated into the spinal cord. However, no proviral DNA was amplified in any neuronal cells, including neurons and glial cells. This indicates that the demyelination of the spinal cord by HTLV-I as a result of viral infection of oligodendrocytes or neuronal cells is unlikely. The T cell receptor V beta gene sequence from lymphocytes in the spinal cord lesions taken from the same HAM/TSP autopsy cases revealed unique and restricted CDR3 motifs, CASSLXG(G) (one-letter amino acid. X is any amino acid), CASSPT(G), and CASSGRL which are similar to those described in T cells from brain lesions of multiple sclerosis (MS) and in a rat T cell clone derived from experimental allergic encephalomyelitis (EAE) lesions. The present results suggest that T cells containing restricted V beta CDR3 motifs, which are also found in MS and EAE, become activated upon HTLV-I infection and infiltrate into the spinal cord lesions of HAM/TSP patients.
确定人类I型嗜T淋巴细胞病毒（HTLV-I）前病毒DNA在中枢神经系统（CNS）中的定位对于理解HTLV-I相关脊髓病（HAM）/热带痉挛性截瘫（TSP）的发病机制至关重要。我们开发了一种灵敏的检测方法，称为两步聚合酶链反应（PCR）原位杂交，该方法使我们能够在HAM/TSP患者石蜡包埋的脊髓组织切片中检测HTLV-I前病毒DNA。仅在浸润到脊髓中的淋巴细胞细胞核中检测到HTLV-I前病毒DNA。然而，在任何神经元细胞（包括神经元和神经胶质细胞）中均未扩增出前病毒DNA。这表明由于少突胶质细胞或神经元细胞的病毒感染，HTLV-I导致脊髓脱髓鞘的可能性不大。从同一HAM/TSP尸检病例的脊髓病变中的淋巴细胞获得的T细胞受体Vβ基因序列显示出独特且受限的CDR3基序，即CASSLXG(G)（单字母氨基酸，X为任意氨基酸）、CASSPT(G)和CASSGRL，这些基序与在多发性硬化症（MS）脑病变的T细胞以及源自实验性自身免疫性脑脊髓炎（EAE）病变的大鼠T细胞克隆中所描述的基序相似。目前的结果表明，含有受限Vβ CDR3基序的T细胞（在MS和EAE中也有发现）在HTLV-I感染后被激活，并浸润到HAM/TSP患者的脊髓病变中。
